3,4-METHYLENEDIOXYAMPHETAMINE Latest Revision: August 16, 2005 1. SYNONYMS CFR: 3,4-Methylenedioxyamphetamine CAS #: Base: 4764-17-4 Hydrochloride: 6292-91-7 Other Names: 3,4-Methylenedioxy-α-methyl-β-phenethylamine 3,4-Methylenedioxyphenylisopropylamine alpha-Methyl-1,3-benzodioxole-5-ethanamine alpha-Methyl-1,3-benzodioxole-5-ethanamine Tenamfetamine Adam Love EA-1299 MDA 2. CHEMICAL AND PHYSICAL DATA 2.1. CHEMICAL DATA Form Chemical Formula Molecular Weight Melting Point (°C) Base C10H13NO2 179.2 Oil Hydrochloride C10H13NO2·HCl 215.7 187-88 2.2. SOLUBILITY Form A C E H M W Base *** S S S S I Hydrochloride SS S I *** S S Sulfate *** I I I S *** A = acetone, C = chloroform, E = ether, H = hexane, M = methanol and W = water, VS = very soluble, FS = freely soluble, S = soluble, PS = sparingly soluble, SS = slightly soluble, VSS = very slightly soluble and I = insoluble 3. SCREENING TECHNIQUES 3.1. COLOR TESTS REAGENT COLOR PRODUCED Marquis Purple to black 3.2. THIN LAYER CHROMATOGRAPHY Visualization Acidified potassium permanganate solution Relative Rf System System COMPOUND TLC 5 TLC 6 3,4-methylenedioxyethylamphetamine 1.1 1.2 3,4-methylenedioxyamphetamine 1.0 1.0 3,4-methylenedioxydimethylamphetamine 0.9 1.2 3,4-methylenedioxymethamphetamine 0.8 0.6 3.3. GAS CHROMATOGRAPHY Method MDA-GCS1 Instrument: Gas Chromatograph operated in split mode with FID Column: 5% phenyl/95% methyl silicone 30 m x 0.25 mm x 0.25 µm film thickness Carrier gas: Hydrogen at 2.0 mL/min Temperatures: Injector: 260°C Detector: 270°C Oven program: 1) 90°C initial temperature for 1.0 min 2) Ramp to 295°C at 30°/min 3) Hold final temperature for 2.6 min Injection Parameters: Split Ratio = 50:1, 1 µL injected Samples are to be dissolved in 4:1 chloroform: methanol and filtered. COMPOUND RRT COMPOUND RRT amphetamine 0.61 acetaminophen 1.24 methamphetamine 0.67 caffeine 1.34 safrole 0.80 methyl stearate 1.49 C14 0.89 cocaine 1.61 MDA 1.0 (4.17 min) methyl eicosanoate 1.62 MDMA 1.05 tetraphenylethylene 1.78 MDEA 1.09 heroin 1.89 MMDA 1.12 Method MDA-GCS2 Instrument: Gas Chromatograph operated in split mode with FID Column: 50% phenyl/50% methyl silicone 30 m x 0.25 mm x 0.25 µm film thickness Carrier gas: Hydrogen at 2.0 mL/min Temperatures: Injector: 260°C Detector: 270°C Oven program: 1) 90oC initial temperature for 1.0 min 2) Ramp to 295°C at 30°/min 3) Hold final temperature for 2.6 min Injection Parameters: Split Ratio = 50:1, 1 µL injected Samples are to be dissolved in 4:1 chloroform: methanol and filtered. COMPOUND RRT COMPOUND RRT amphetamine 0.62 acetaminophen 1.31 methamphetamine 0.66 caffeine 1.38 safrole 0.80 methyl stearate 1.30 C14 0.69 cocaine 1.58 MDA 1.0 (4.84 min) methyl eicosanoate 1.41 MDMA 1.03 tetraphenylethylene 1.77 MDEA 1.05 heroin 2.03 MMDA 1.07 Method MDA-GCS3 Instrument: Gas Chromatograph operated in split mode with FID Column: 5% phenyl/95% methyl silicone 5 m x 0.25 mm x 0.25µm film thickness Carrier gas: Hydrogen: 1) Initial pressure of 2.5 psi for 0.10 min 2) Ramp to 5.0 psi at 3.41 psi/min 3) Hold final pressure for 0.77 min Temperature: Injector: 260°C Detector: 270°C Oven program: 1) 90°C initial temperature 2) Ramp to 100°C at 10°C/min 3) Hold final temperature for 0.60 min Injection Parameters: Split Ratio = 50:1, 1 µL injected EZFlash Parameters: 1) Time 0 sec = 100°C 2) Time 6 sec = 110°C 3) Time 50 sec = 310°C 4) Time 96 sec = 310°C Samples are to be dissolved in 4:1 chloroform:methanol and filtered. COMPOUND RRT COMPOUND RRT amphetamine 0.73 acetaminophen 1.19 methamphetamine 0.77 caffeine 1.27 safrole 0.85 methyl stearate 1.28 C14 0.92 cocaine 1.50 MDA 1.0 (4.79 min) methyl eicosanoate 1.40 MDMA 1.04 tetraphenylethylene 1.69 MDEA 1.07 heroin 1.88 MMDA 1.08 Method MDA-GCS4 Instrument: Gas Chromatograph operated in split mode with FID Column: 1% phenyl/99% methyl silicone 30 m x 0.25 mm x 0.25 µm film thickness Carrier gas: Hydrogen at 2.0 mL/min Temperature: Injector: 260°C Detector: 270°C Oven program: 1) 90°C initial temperature for 1.0 min 2) Ramp to 295°C at 30°/min 3) Hold final temperature for 2.6 min Injection Parameters: Split Ratio = 50:1, 1 µL injected COMPOUND RRT COMPOUND RRT amphetamine 0.56 acetaminophen 1.27 methamphetamine 1.75 caffeine 1.36 safrole 0.77 methyl stearate 1.63 C14 0.97 cocaine 1.70 MDA 1.0 (4.1 min) methyl eicosanoate 1.78 MDMA 1.06 tetraphenylethylene 1.89 MDEA 1.12 heroin 2.01 MMDA 1.15 4. SEPARATION TECHNIQUES 5. QUANTITATIVE PROCEDURE 5.1. GAS CHROMATOGRAPHY Method MDA-GCQ1 Internal Standard Stock Solution: 1.0 mg/mL n-butylamphetamine HCl in chloroform. Standard Solution Preparation: Prepare a standard solution of MDA HCl at 0.4 mg/mL in chloroform. Sample Preparation: Accurately weigh out 50 mg of sample into a 25 mL volumetric flask and dilute with water to the mark. Take a 2 mL aliquot of the solution, add 3 mL of I.S. and 2-3 mL of 0.5 M KOH. Shake. Separate layers and extract twice more with 2-3 mL each time of chloroform. Combine fractions – dry over sodium sulfate and dilute to total volume of 10 mL. Treat the standard in the same fashion as the sample prior to analysis. Instrument: Gas Chromatograph operated in split mode with FID Column: DB-1, 30 m x 0.25 mm x 0.25 m film thickness Carrier gas: Hydrogen at 1.3 mL/min Make-Up gas: Nitrogen at 40.0 mL/min Temperatures: Injector: 230°C Detector: 280°C Oven Program: 1) 150°C initial temperature 2 min 2) Ramp to 190°C at 5°C/min 3) Hold final temperature for 1.0 min Injection Parameters: Split Ratio = 20:1, 1µL injected Typical Retention Time: 3,4-MDA: 2.13 min N-Butylamphetamine HCl: 1.89 min Linear Range: 0.06 mg/mL to 2.0 mg/mL Repeatability: RSD less than 2.0% Correlation Coefficient: 0.9998 Accuracy Error less than 5% COMPOUND RRT COMPOUND RRT COMPOUND RRT methylbenzylamine 0.40 salicylamide 0.89 caffeine 2.14 P2P 0.44 methyl paraben 0.89 antipyrine 2.30 amphetamine 0.45 penicillin 0.90 benzphetamine 2.35 nicotinic acid 0.48 phenmetrazine 0.93 diphenhydramine 2.46 phentermine 0.48 MDP-2-P 0.97 aminopyrine 2.60 methamphetamine 0.51 phendimetrazine 0.99 doxylamine 2.66 COMPOUND RRT COMPOUND RRT COMPOUND RRT fenfluramine 0.55 MDA 1.0 (2.13 min) phthalic acid 2.82 ethylamphetamine 0.56 MDMA 1.15 palmitic acid 2.82 dimethylamphetamine 0.58 aminorex 1.21 dipyrone 2.89 safrole 0.63 methyl aminorex 1.21 procaine 3.06 salicylic acid 0.64 MDEA 1.30 eicosane 3.07 cathine 0.68 ibuprofen 1.43 dextromethorphan 3.59 phenylpropanolamine 0.68 MBDB 1.45 strychnine 3.65 methcathinone 0.69 hexadecane 1.47 amitriptyline 3.84 nicotinamide 0.71 guaifenesin 1.50 scopolamine 4.27 chlorpheniramine 0.75 acetaminophen 1.55 tetracosane 4.58 chloroephedrine 0.76 MMDA 1.59 chlordiazepoxide 4.96 pseudoephedrine 0.76 phenacetin 1.62 quinine 6.02 ephedrine 0.78 chloromdma 1.71 butylamphetamine 0.89 methylphenidate 1.87 5.2. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY Method MDA-LCQ1 Standard Solution Preparation: Accurately weigh and prepare a standard solution of MDA HCl at approximately 1.0 mg/mL using water. Filter with a 0.45-µm filter. Sample Preparation: Accurately weigh an amount of sample into a volumetric flask and dilute with water. If necessary dilute the sample so the final concentration approximates the standard concentration or falls within the linear range. Filter sample with a 0.45-µm filter. Instrument: High performance liquid chromatograph equipped with diode array Column: 5 µm ODS, 4.0 mm x 125 mm Detector: UV, 210 nm Flow: 1.0 mL/min Injection Volume: 1 µL Buffer: 4000 mL distilled water, 30 mL phosphoric acid, 10 g sodium hydroxide and 8.0 mL hexylamine at pH 2.5 Mobile Phase: Buffer: acetonitrile 85:15 with gradient to 60:40 over 6 min Typical Retention Time: 3,4-MDA: 1.63 min Linear Range: 0.1338 - 4.28 mg/mL Repeatability: RSD less than 0.2% Correlation Coefficient: 0.999 Accuracy: Error less than 5% COMPOUND RRT Nicotinamide 0.62 MDA 1.00 MDMA 1.16 MDEA 1.41 Method MDA-LCQ2 Standard Solution Preparation: Prepare a standard solution of MDA HCl at approximately 0.5 mg/mL using water, buffer, or methanol. Sample Preparation: Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the final MDA HCl concentration is approximately that of the standard solution. Instrument: High performance liquid chromatograph equipped with diode array Column: 5 Phenomenex Luna, 150 mm x 3.2 mm at 35 C Detector: UV, 210 nm Flow: 1.0 mL/min Injection Volume: 5 µL Buffer: 4000 mL distilled water, 22.5 mL phosphoric acid, adjust to pH 2.2-2.3 with triethanolamine (approx. 22 mL) Mobile Phase: Buffer:acetonitrile 90:10 Typical Retention Time: 3,4-MDA: 9.8 min Linear Range: 0.05 mg/mL - 2 mg/mL Repeatability: RSD less than 0.5% Correlation Coefficient: 0.9999 Accuracy: Error less than 5% COMPOUND RRT COMPOUND RRT phenylpropanolamine 0.65 methyl aminorex 1.09 methyl benzylamine 0.67 phentermine 1.10 cathine 0.68 chloroephedrine 1.11 doxylamine 0.74 ethylamphetamine 1.13 dipyrone 0.75 MDP-2-P 1.13 ephedrine 0.77 lidocaine 1.15 methcathinone 0.79 caffeine 1.16 pseudoephedrine 0.79 MDEA 1.22 amphetamine 0.90 phenyl-2-naphthalene 1.22 theophylline 0.94 phthalic acid 1.22 methapyrilene 0.96 strychnine 1.23 phenmetrazine 0.96 MMDA 1.26 methamphetamine 0.98 MBDB 1.34 phendimetrazine 0.98 salicylamide 1.37 scopolamine 0.99 acetaminophen 1.39 COMPOUND RRT COMPOUND RRT MDA 1.00 (9.8 min) chloro-mdma 1.47 quinine 1.03 methylphenidate 1.48 saccharin 1.03 glycerol glycolate 1.59 dimethylamphetamine 1.04 chlorpheniramine 1.67 MDMA 1.08 aspirin 1.95 chlordiazepoxide 2.01 5.3.